scholarly journals Hypoxia-inducible factor 1α induces osteo/odontoblast differentiation of human dental pulp stem cells via Wnt/β-catenin transcriptional cofactor BCL9

2022 ◽  
Vol 12 (1) ◽  
Author(s):  
Shion Orikasa ◽  
Nobuyuki Kawashima ◽  
Kento Tazawa ◽  
Kentaro Hashimoto ◽  
Keisuke Sunada-Nara ◽  
...  
Keyword(s):  
Stem Cells ◽  
Dental Pulp ◽  
Target Genes ◽  
Hypoxia Inducible Factor ◽  
Dental Pulp Stem Cells ◽  
Pulp Tissue ◽  
Odontoblast Differentiation ◽  
Hypoxia Inducible Factor 1Α ◽  
Hypoxic Conditions

AbstractAccelerated dental pulp mineralization is a common complication in avulsed/luxated teeth, although the mechanisms underlying this remain unclear. We hypothesized that hypoxia due to vascular severance may induce osteo/odontoblast differentiation of dental pulp stem cells (DPSCs). This study examined the role of B-cell CLL/lymphoma 9 (BCL9), which is downstream of hypoxia-inducible factor 1α (HIF1α) and a Wnt/β-catenin transcriptional cofactor, in the osteo/odontoblastic differentiation of human DPSCs (hDPSCs) under hypoxic conditions. hDPSCs were isolated from extracted healthy wisdom teeth. Hypoxic conditions and HIF1α overexpression induced significant upregulation of mRNAs for osteo/odontoblast markers (RUNX2, ALP, OC), BCL9, and Wnt/β-catenin signaling target genes (AXIN2, TCF1) in hDPSCs. Overexpression and suppression of BCL9 in hDPSCs up- and downregulated, respectively, the mRNAs for AXIN2, TCF1, and the osteo/odontoblast markers. Hypoxic-cultured mouse pulp tissue explants showed the promotion of HIF1α, BCL9, and β-catenin expression and BCL9-β-catenin co-localization. In addition, BCL9 formed a complex with β-catenin in hDPSCs in vitro. This study demonstrated that hypoxia/HIF1α-induced osteo/odontoblast differentiation of hDPSCs was partially dependent on Wnt/β-catenin signaling, where BCL9 acted as a key mediator between HIF1α and Wnt/β-catenin signaling. These findings may reveal part of the mechanisms of dental pulp mineralization after traumatic dental injury.

scholarly journals Taxifolin Protects Dental Pulp Stem Cells under Hypoxia and Inflammation Conditions

2021 ◽  
Vol 30 ◽  
pp. 096368972110344
Author(s):  
Xiaohui Fu ◽  
Yimiao Feng ◽  
Bingyi Shao ◽  
Yanzhen Zhang
Keyword(s):  
Stem Cells ◽  
Dental Pulp ◽  
Annexin V ◽  
Protective Role ◽  
Dental Pulp Stem Cells ◽  
Pulp Tissue ◽  
Inflammatory Conditions ◽  
Human Dental Pulp ◽  
Unique Source

Background: Dental pulp stem cells (DPSCs) are a unique source for future clinical application in dentistry such as periodontology or endodontics. However, DPSCs are prone to apoptosis under abnormal conditions. Taxifolin is a natural flavonoid and possesses many pharmacological activities including anti-hypoxic and anti-inflammatory. We aimed to elucidate the mechanisms of taxifolin protects DPSC under hypoxia and inflammatory conditions. Methods: DPSCs from human dental pulp tissue was purchased from Lonza (cat. no. PT-5025. Basel, Switzerland)) and identified by DPSC’s biomarkers. DPSC differentiation in vitro following the manufacturers’ instructions. ARS staining and Oil red staining verify the efficiency of differentiation in vitro after 2 weeks. The changes of various genes and proteins were identified by Q-PCR and western-blot, respectively. Cell viability was determined by the CCK-8 method, while apoptosis was determined by Annexin V/PI staining. Results: DPSC differentiation in vitro shows that hypoxia and TNF-α synergistically inhibit the survival and osteogenesis of DPSCs. A final concentration of 10 μM Taxifolin can significantly reduce the apoptosis of DPSCs under inflammation and hypoxia conditions. Taxifolin substantially increases carbonic anhydrase IX (CA9) expression but not HIF1a, and inhibitions of CA9 expression nullify the protective role of taxifolin under hypoxia and inflammatory condition. Conclusion: Taxifolin significantly increased the expression of CA9 when it inhibits DPSC apoptosis and taxifolin synergistically to protect DPSCs against apoptosis with CA9 under hypoxia and inflammatory conditions. Taxifolin can be used as a potential drug for clinical treatment of DPSC-related diseases.

scholarly journals VEGFR1 primes a unique cohort of dental pulp stem cells for vasculogenic differentiation

2021 ◽  
Vol 41 ◽  
pp. 332-344
Author(s):  
MT Bergamo ◽  
◽  
Z Zhang ◽  
TM Oliveira ◽  
JE Nör
Keyword(s):  
Stem Cells ◽  
Dental Pulp ◽  
Vascular Endothelial Cells ◽  
Dental Pulp Stem Cells ◽  
Deciduous Teeth ◽  
Microvascular Endothelial Cell ◽  
Vascular Endothelial ◽  
Pulp Tissue ◽  
Immunodeficient Mice

Dental pulp stem cells (DPSCs) constitute a unique group of cells endowed with multipotency, self-renewal, and capacity to regenerate the dental pulp tissue. While much has been learned about these cells in recent years, it is still unclear if each DPSC is multipotent or if unique sub-populations of DPSCs are “primed” to undergo specific differentiation paths. The purpose of the present study was to define whether a sub-population of DPSCs was uniquely primed to undergo vasculogenic differentiation. Permanent-tooth DPSCs or stem cells from human exfoliated deciduous teeth (SHED) were flow-sorted for vascular endothelial growth factor receptor 1 (VEGFR1) and exposed to vasculogenic differentiation medium, i.e., Microvascular-Endothelial-Cell-Growth-Medium-2-BulletKit™ supplemented with 50 ng/mL rhVEGF165 in the presence of 0 or 25 μg/mL anti-human VEGF antibody (bevacizumab; Genentech). In addition, sorted SHED (i.e., VEGFR1high or VEGFR1low) were seeded in biodegradable scaffolds and transplanted into the subcutaneous space of immunodeficient mice. Despite proliferating at a similar rate, VEGFR1high generated more in vitro sprouts than VEGFR1low cells (p < 0.05). Blockade of VEGF signaling with bevacizumab inhibited VEGFR1high-derived sprouts, demonstrating specificity of responses. Similarly, VEGFR1high SHED generated more blood vessels when transplanted into murine hosts than VEGFR1low cells (p < 0.05). Collectively, these data demonstrated that DPSCs contain a unique sub-population of cells defined by high VEGFR1 expression that are primed to differentiate into vascular endothelial cells. These data raise the possibility of purifying stem cells with high vasculogenic potential for regeneration of vascularized tissues or for vascular engineering in the treatment of ischemic conditions.

Biological Effects of Provisional Resin Materials on Human Dental Pulp Stem Cells

2017 ◽  
Vol 42 (2) ◽  
pp. E81-E92 ◽  
Author(s):  
S-K Jun ◽  
C Mahapatra ◽  
H-H Lee ◽  
H-W Kim ◽  
J-H Lee
Keyword(s):  
Stem Cells ◽  
Proinflammatory Cytokines ◽  
Repeated Measures ◽  
Dental Pulp ◽  
Culture Media ◽  
Biological Effects ◽  
Dental Pulp Stem Cells ◽  
Pulp Tissue ◽  
Human Dental Pulp

SUMMARY Objectives: This study investigated the in vitro cytotoxicity as well as the proinflammatory cytokine expression of provisional resin materials on primary cultured human dental pulp stem cells (hDPSCs). Methods: Five commercially available provisional resin materials were chosen (SNAP [SN], Luxatemp [LT], Jet [JE], Revotek LC [RL], and Vipi block [VB]). Eluates that were either polymerizing or already set were added to hDPSCs under serially diluted conditions divided into three different setting times (25% set, 50% set, and 100% set) and incubated for 24 hours with 2× concentrated culture media. Cell cytotoxicity tests were performed by LDH assay and live and dead confocal microscope images. The expression of proinflammatory cytokines in SN and VB was measured using cytokine antibody arrays. Data were analyzed using repeated measures analysis of variance (ANOVA) or ANOVA followed by the Tukey post hoc test at a significance level of p&lt;0.05. Results: Cytotoxicity greater than 30% was observed in the 50% diluted culture in SN, LT, and JE in the already set stage (p&lt;0.05), while it was detected in SN and LT in early or intermediate stage samples. The cytotoxicity of SN, JE, and LT was greater with eluates from the polymerizing phase compared to that from already set samples (p&lt;0.05), as observed by live and dead images. On the other hand, RL and VB did not exhibit cytotoxicity greater than 30%. Proinflammatory cytokines were not detected in 12.5% diluted culture with eluates from VB and early set stage SN. Conclusions: The eluates from chemical-activated provisional resin materials during polymerization (SN, LT, and JE) were cytotoxic to hDPSCs and may adversely affect pulp tissue.

scholarly journals Importance of In Vitro Microenvironment on Differentiation and Translational Applications of Dental Pulp Stem Cells

2021 ◽  
Vol 7 (1) ◽  
Author(s):  
Li Q ◽  
Keyword(s):  
Stem Cells ◽  
Dental Pulp ◽  
Adult Stem Cells ◽  
Nerve Repair ◽  
Dental Pulp Stem Cells ◽  
Pulp Tissue ◽  
Cell Microenvironment ◽  
Dental Pulp Tissue ◽  
Multipotent Differentiation

Dental Pulp Stem Cells (DPSCs) are adult stem cells found in dental pulp tissue, and possess the capacity for self-renewal and the potential for multipotent differentiation. DPSCs depend on regulating in vitro microenvironment, and can readily differentiate into osteoblasts, odontoblasts, neurocytes, adipocytes, chondrocytes, myocytes, fibrocytes and many others. In addition, DPSCs play a crucial role in tooth regeneration, bone and nerve repair in current studies of regenerative medicine and tissue engineering. Among them, the influences of stem cell microenvironment or niche on the biological activity of DPSCs are critical and hamper its progress. Herein, we review the influence of culture condition, tissue source, growth factor requirements, and cellular organizational scaffolds and how these features influence the biological characteristics and translational research of DPSCs.

Isolation and characterization of human dental pulp stem cells (DPSCs) and stem cells from human exfoliated deciduous teeth (SHED) – an in vitro study

2018 ◽  
Vol 10 (2) ◽  
pp. 9-18
Author(s):  
Dr. Vinay Rao ◽  
Dr. Roopa R Nadig ◽  
Dr.Ramanand Nadig ◽  
Dr.Karthik .J ◽  
Dr. Veena Pai S
Keyword(s):  
Stem Cells ◽  
Dental Pulp ◽  
Surface Characterization ◽  
Dental Pulp Stem Cells ◽  
In Vivo Studies ◽  
Deciduous Teeth ◽  
Pulp Tissue ◽  
Isolation And Characterization

AIMS AND OBJECTIVES: 1. Isolation and growth of dental pulp stem cells (DPSCs) and stem cells from exfoliated human deciduous teeth (SHED). 2. Characterization of dental pulp stem cells (DPSCs) and stem cells from exfoliated human deciduous teeth (SHED). METHODS: The pulp tissue was digested in collagenase and cultured in DMEM Dulbecco’s Modified Eagle’s Media). The stem cells were identified and isolated. Surface characterization of cells was done with the help of flow cytometer using a panel of various surface markers. An immuno cytochemistry analysis was done to see the expression of proteins in the cells. RESULTS: Identification of cells was done with the help of a phase contrast microscope. Flow cytometry analyses for various CD markers showed similar results for both DPSCs and SHED. The cells showed positive expression for pluripotent markers, ectodermal markers and mesodermal markers. CONCLUSION: The study demonstrated that stem cells existed in human deciduous and permanent pulp tissue. The stem cells present in deciduous permanent pulp tissue can be isolated, cultivated and expanded in vitro. Both DPSCs and SHED show almost a similar expression pattern profile for variety of antigens tested. Further studies should include analysis of diverse cell populations to elucidate their potential to differentiate into various cell types followed by in vivo studies in animals and humans.

Stem Cells Derived from Human Exfoliated Deciduous teeth [shed] in Neuronal Disorders: A Review

2020 ◽  
Vol 16 ◽  
Author(s):  
Minu Anoop ◽  
Indrani Datta
Keyword(s):  
Stem Cells ◽  
Neurodegenerative Diseases ◽  
Dental Pulp ◽  
Therapeutic Potential ◽  
Permanent Teeth ◽  
Deciduous Teeth ◽  
Proliferative Potential ◽  
Pulp Tissue ◽  
Human Exfoliated Deciduous Teeth

: Most conventional treatments for neurodegenerative diseases fail due to their focus on neuroprotection rather than neurorestoration. Stem cell‐based therapies are becoming a potential treatment option for neurodegenerative diseases as they can home in, engraft, differentiate and produce factors for CNS recovery. Stem cells derived from human dental pulp tissue differ from other sources of mesenchymal stem cells due to their embryonic neural crest origin and neurotrophic property. These include both dental pulp stem cells [DPSCs] from dental pulp tissues of human permanent teeth and stem cells from human exfoliated deciduous teeth [SHED]. SHED offer many advantages over other types of MSCs such as good proliferative potential, minimal invasive procurement, neuronal differentiation and neurotrophic capacity, and negligible ethical concerns. The therapeutic potential of SHED is attributed to the paracrine action of extracellularly released secreted factors, specifically the secretome, of which exosomes is a key component. SHED and its conditioned media can be effective in neurodegeneration through multiple mechanisms, including cell replacement, paracrine effects, angiogenesis, synaptogenesis, immunomodulation, and apoptosis inhibition, and SHED exosomes offer an ideal refined bed-to-bench formulation in neurodegenerative disorders. However, in spite of these advantages, there are still some limitations of SHED exosome therapy, such as the effectiveness of long-term storage of SHED and their exosomes, the development of a robust GMP-grade manufacturing protocol, optimization of the route of administration, and evaluation of the efficacy and safety in humans. In this review, we have addressed the isolation, collection and properties of SHED along with its therapeutic potential on in vitro and in vivo neuronal disorder models as evident from the published literature.

scholarly journals Enhanced Osteogenesis of Dental Pulp Stem Cells In Vitro Induced by Chitosan–PEG-Incorporated Calcium Phosphate Cement

Polymers ◽  
2021 ◽  
Vol 13 (14) ◽  
pp. 2252
Author(s):  
Jae Eun Kim ◽  
Sangbae Park ◽  
Woong-Sup Lee ◽  
Jinsub Han ◽  
Jae Woon Lim ◽  
...  
Keyword(s):  
Stem Cells ◽  
Calcium Phosphate ◽  
Zeta Potential ◽  
Mechanical Strength ◽  
Calcium Phosphate Cement ◽  
Dental Pulp ◽  
Dental Pulp Stem Cells ◽  
Ion Trapping ◽  
Alizarin Red

The use of bone graft materials is required for the treatment of bone defects damaged beyond the critical defect; therefore, injectable calcium phosphate cement (CPC) is actively used after surgery. The application of various polymers to improve injectability, mechanical strength, and biological function of injection-type CPC is encouraged. We previously developed a chitosan–PEG conjugate (CS/PEG) by a sulfur (VI) fluoride exchange reaction, and the resulting chitosan derivative showed high solubility at a neutral pH. We have demonstrated the CPC incorporated with a poly (ethylene glycol) (PEG)-grafted chitosan (CS/PEG) and developed CS/PEG CPC. The characterization of CS/PEG CPC was conducted using Fourier transform infrared spectroscopy (FT-IR) and X-ray diffraction (XRD). The initial properties of CS/PEG CPCs, such as the pH, porosity, mechanical strength, zeta potential, and in vitro biocompatibility using the WST-1 assay, were also investigated. Moreover, osteocompatibility of CS/PEG CPCs was carried out via Alizarin Red S staining, immunocytochemistry, and Western blot analysis. CS/PEG CPC has enhanced mechanical strength compared to CPC, and the cohesion test also demonstrated in vivo stability. Furthermore, we determined whether CS/PEG CPC is a suitable candidate for promoting the osteogenic ability of Dental Pulp Stem Cells (DPSC). The elution of CS/PEG CPC entraps more calcium ion than CPC, as confirmed through the zeta potential test. Accordingly, the ion trapping effect of CS/PEG is considered to have played a role in promoting osteogenic differentiation of DPSCs. The results strongly suggested that CS/PEG could be used as suitable additives for improving osteogenic induction of bone substitute materials.

scholarly journals Odontoblast-Like Cells Differentiated from Dental Pulp Stem Cells Retain Their Phenotype after Subcultivation

2018 ◽  
Vol 2018 ◽  
pp. 1-12 ◽  
Author(s):  
Paula A. Baldión ◽  
Myriam L. Velandia-Romero ◽  
Jaime E. Castellanos
Keyword(s):  
Stem Cells ◽  
Dental Pulp ◽  
Matrix Protein ◽  
Dental Materials ◽  
Dental Pulp Stem Cells ◽  
Cell Physiology ◽  
Pulp Tissue ◽  
Alizarin Red ◽  
Dentin Matrix Protein ◽  
Mineral Deposition

Odontoblasts, the main cell type in teeth pulp tissue, are not cultivable and they are responsible for the first line of response after dental restauration. Studies on dental materials cytotoxicity and odontoblast cells physiology require large quantity of homogenous cells retaining most of the phenotype characteristics. Odontoblast-like cells (OLC) were differentiated from human dental pulp stem cells using differentiation medium (containing TGF-β1), and OLC expanded after trypsinization (EXP-21) were evaluated and compared. Despite a slower cell growth curve, EXP-21 cells express similarly the odontoblast markers dentinal sialophosphoprotein and dentin matrix protein-1 concomitantly with RUNX2 transcripts and low alkaline phosphatase activity as expected. Both OLC and EXP-21 cells showed similar mineral deposition activity evidenced by alizarin red and von Kossa staining. These results pointed out minor changes in phenotype of subcultured EXP-21 regarding the primarily differentiated OLC, making the subcultivation of these cells a useful strategy to obtain odontoblasts for biocompatibility or cell physiology studies in dentistry.

P-EP013. Superior migration propensity of dental pulp stem cells in in vitro and in vivo models of excitotoxic neurodegeneration

2021 ◽  
Vol 132 (8) ◽  
pp. e82-e83
Author(s):  
Sivapriya Senthilkumar ◽  
Chaitra Venugopal ◽  
K. Shobha ◽  
Bindu M. Kutty ◽  
Anandh Dhanushkodi
Keyword(s):  
Stem Cells ◽  
Dental Pulp ◽  
Dental Pulp Stem Cells ◽  
In Vivo Models ◽  
Migration Propensity
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